362 2300 x 144
>-Original Message-
>From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On Behalf Of
>Benini, Stefano
>Sent: Wednesday, July 11, 2007 7:17 AM
>To: CCP4BB@JISCMAIL.AC.UK
>Subject: Re: [ccp4bb] Help with reducing crystal mosaicity
>
>Dear Mary,
>
rk, California.
From: CCP4 bulletin board on behalf of Kris Tesh
Sent: Wed 7/11/2007 8:35 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Help with reducing crystal mosaicity
At the risk of sounding too commercial, here are some suggestions:
1. See if you can eit
Sent: Wednesday, July 11, 2007 7:17 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Help with reducing crystal mosaicity
Dear Mary,
the problem encountered with cryoprotectans is the change in the solution
surrounding your crystal as they may not be present in your crystallisation
conditions or
erley Park
-Original Message-
From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] Behalf Of
Patrick Shaw Stewart
Sent: 11 July 2007 12:10
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Help with reducing crystal mosaicity
Just a thought, Mary - going back to your original question abo
age-
> From: CCP4 bulletin board [mailto:[EMAIL PROTECTED] On Behalf Of Mary
> Fitzgerald
> Sent: 09 July 2007 23:05
> To: CCP4BB@JISCMAIL.AC.UK
> Subject: [ccp4bb] Help with reducing crystal mosaicity
>
> Help please!
>
> I'm looking for some new ideas. I have crystal
ology
Department of Biochemistry
Henry Wellcome Building
University of Leicester
LE1 9HN
0116 229 7097
From: CCP4 bulletin board on behalf of mesters
Sent: Wed 11/07/2007 08:46
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Help with reducing crystal mosaicity
snipped
We used to use FTS-cooler for stable
temperatures in Laue-experiments on
protein crystals in capillaries.
Check out BioCARS at the APS for this device.
http://cars9.uchicago.edu/biocars/pages/timeresolved.html
It has a big opening for constant air flow
and temperature along the capillary and
it c
Matthew,
I have not had any problem with our Cryojet XL (90 - 300 Kelvin), it is
very stable at sub-zero temperatures, I checked it over a longer period
using a highly sensitive electronic device. Also, you can buy the
Cryojet HT that can be operated from 90 – 490 Kelvin!
The heaters will be
CCP4 bulletin board wrote on 07/10/2007 02:07:37
PM:
> Jeroen brings up a good point. Back in the old days, around 5 B. C.
> (Before Cryo), we would use a chilled air generator to blow a stream
> of cold air along the capillary axis to keep the crystals just above
> their freezing point--it made
Jeroen brings up a good point. Back in the old days, around 5 B. C.
(Before Cryo), we would use a chilled air generator to blow a stream
of cold air along the capillary axis to keep the crystals just above
their freezing point--it made a huge difference in crystal lifetime.
I recall a coll
egrees! At 277 K, the crystals only last for a few
images and freezing did not work (for the buffer mentioned before).
- J. -
> -Original Message-
> From: Mary Fitzgerald <[EMAIL PROTECTED]>
> Date: Mon, 9 Jul 2007 18:05:10
> To:CCP4BB@JISCMAIL.AC.UK
> Sub
the buffer mentioned before).
- J. -
> -Original Message-
> From: Mary Fitzgerald <[EMAIL PROTECTED]>
> Date: Mon, 9 Jul 2007 18:05:10
> To:CCP4BB@JISCMAIL.AC.UK
> Subject: [ccp4bb] Help with reducing crystal mosaicity
>
> Help please!
>
> I'm l
re).
- J. -
-Original Message-
From: Mary Fitzgerald <[EMAIL PROTECTED]>
Date: Mon, 9 Jul 2007 18:05:10
To:CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Help with reducing crystal mosaicity
Help please!
I'm looking for some new ideas. I have crystals that come out of
Hi Mary,
not sure if you received this suggestion already.
Despite the low resolution diffraction 4-5 A as you report, try mounting
smaller crystals and see how well they perform. Very often our crystals
had a better freezing experience when they were small, even if
sufficient cryo protectant
On Tuesday 10 July 2007 09:16, Mary Fitzgerald wrote:
> If that doesn't work maybe, I'll try seeding at lower MPD
> concentrations or pressure freezing.
>
> Thanks again,
> Mary
With such a high MPD condition I would probably first try micro-seeding at
25-35% MPD before doing anything else. It so
Wow, thanks. I'm going to try to answer most of the questions I've
received in one message as I'm overwhelmed by the quick multitude of
responses.
As I stated earlier, I haven't collected any room temperature data,
yet. So, I don't know the unfrozen mosaicity. It is very possible
that the cry
Hi Mary,
What final percent MPD do you have prior to flashcooling? Karolin Luger (and
perhaps others) found
that the final percent MPD had a significant effect on crystal mosaicity and
final diffraction
limit for nucleosome crystals.
For example, if the crystals are left in mother liquor con
Here's a method that has had some success in reducing mosaicity, either
with or without cryoprotectant:
Chae Un Kim, Raphael Kapfer and Sol M. Gruner (2005), High Pressure
Cooling of Protein Crystals without Cryoprotectants, ActaCryst. D61,
881-890
This method may be available at a synchrotron ne
What is the mosaicity of the unfrozen crystal?
Jim
-Original Message-
From: Mary Fitzgerald <[EMAIL PROTECTED]>
Date: Mon, 9 Jul 2007 18:05:10
To:CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] Help with reducing crystal mosaicity
Help please!
I'm looking for some new ide
Help please!
I'm looking for some new ideas. I have crystals that come out of a
sitting drop with a mixture of sodium cacodylate at pH 6.5, magnesium
acetate and MPD for the well solution. The MPD concentration is
sufficient to act as a cryoprotectant. Currently, I directly freeze
these crysta
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